脐血与脐带间充质干细胞的分离、扩增、分化比较

目的比较人脐血、脐带组织间充质干细胞体外分离、扩增与成骨、成脂肪细胞分化的方法与条件．比较相应实验的难易度。方法脐血采用沉降红细胞后密度梯度法及CD34^＋免疫磁珠负选法分离单个核细胞＋10％胎牛血清或Mesencuit^TM培养基培养传代，集落生长细胞向成骨、脂肪细胞定向诱导分化；脐带沿血管灌入胶原酶悬液，获取细胞培养、扩增，细胞呈集落生长后传代，定向成骨、成脂肪细胞分化。结果脐血经沉降红细胞后分离的MNCs，使用Mesencult^TM培养基＋10％胎牛血清培养成功率高，集落细胞能向成骨、成脂肪细胞定向诱导分化。脐带去除血管后灌注可获得贴壁生长的细胞，能扩增形成集落，并向成骨、成脂肪细胞定向分化。结论脐血中可分离出MSCs，并可在体外进行培养扩增及分化，但难度较大。脐带组织存在间充质干细胞，并可在体外进行培养扩增形成集落细胞传代，集落细胞能够向成骨、成脂肪细胞分化，方法较脐血容易。

Umbilical Cord Blood and Umbilical Cord Mesenchymal StemCells Isolated, Proliferated, Differentiation Compared

Objective Comparison of methods and conditions for human umbilical cord blood, umbilical cord tissue-derived mesenchymal stem cells in vitro isolation, amplification and into osteogenic and adipogenic differentiation directly. Method Human umbilical cord blood sedimented red cell with methylcellulose then centrifugated,, or obtained by negative immunodepletion of CD34^＋. These isolated cells were cultured and proliferated clonally in medium which consists of L-DMEM or Meseneult^TM medium（＋ 10% fetal calf serum（FCS） respectively）, colony growth of cells able to differentiate into osteoblast and lipoblast. Umbilical cord blood vessels was poured into the collagenase suspension, we got suspended cells and centrifuged, Finally, the cells were cultured in polystyrene dishes, we obtained single cell-derived colonies, then we differentiated them to osteoblast and lipoblast. Result The cord blood mononuclear cells isolated by sedimented and centrifugated way cultured in Mesencult^TM medium＋10% FCS are most available. From the tissues of umbilical cord we can get adherent cells, it can be proliferated and form single cell-derived colonies, able to differentiated into osteoblast and lipoblast. Conclusion We can isolate MSCs from human umbilical cord blood and proliferate it in vitro, but the more difficult.We can isolate MSCs from the tissues of umbilical cord and proliferate it in vitro, proliferation colonies can differentiate into osteoblasts and adipocytes, the way is easier than the cord blood＇s.